Ectromelia virus(ECTV),a member of the Orthopoxvirus genus,serves as both a causative agent of mousepox and a pivotal surrogate model for studying highly pathogenic orthopoxviruses.Although genomic data on ECTV remain...Ectromelia virus(ECTV),a member of the Orthopoxvirus genus,serves as both a causative agent of mousepox and a pivotal surrogate model for studying highly pathogenic orthopoxviruses.Although genomic data on ECTV remains limited,we report the isolation and characterization of a novel strain,ECTV-C-Tan-GD01,obtained from rodents in Guangdong Province,China.Nanopore sequencing yielded a complete genome(199 annotated genes,including one gene truncated at the C-terminus)with inverted terminal repeats(ITRs)harboring a conserved hairpin structure.Notably,a frameshift-inducing“G”deletion in the EV159 gene resulted in the truncation of a semaphorin-like protein.In vitro assays demonstrated cell-associated viral replication kinetics,with maximum titers achieved earlier in Vero/HeLa cells(72 h)than in BHK-21/CEF cells(84 h).Murine challenge experiments revealed extreme virulence(LD50<1 plaque-forming unit(PFU)via intranasal/footpad routes)and hepatosplenic tropism.Furthermore,ECTV-C-Tan-GD01 exhibited utility in evaluating orthopoxvirus countermeasures:a single dose of vaccinia virus Tiantan(VTT)or non-replicating vaccinia virus Tiantan(NTV)conferred cross-protection,while tecovirimat(ST-246),cidofovir(CDV),and brincidofovir(initially CMX001)significantly reduced viral loads and pathology.This study establishes ECTV-C-Tan-GD01 as a dual-purpose resource for probing orthopoxvirus evolution and advancing therapeutic development.展开更多
Emerging and re-emerging viruses from wild animals have seriously threatened the health of humans and domesticated animals in recent years.Herein,we isolated a new mammalian orthoreovirus(MRV),Pika/MRV/GCCDC7/2019(PMR...Emerging and re-emerging viruses from wild animals have seriously threatened the health of humans and domesticated animals in recent years.Herein,we isolated a new mammalian orthoreovirus(MRV),Pika/MRV/GCCDC7/2019(PMRV-GCCDC7),in the Qinghai-Tibet Plateau wild pika(Ochotona curzoniae).Though the PMRVGCCDC7 shows features of a typical reovirus with ten gene segments arranged in 3:3:4 in length,the virus belongs to an independent evolutionary branch compared to other MRVs based on phylogenetic tree analysis.The results of cellular susceptibility,species tropism,and replication kinetics of PMRV-GCCDC7 indicated the virus could infect four human cell lines(A549,Huh7,HCT,and LoVo)and six non-human cell lines,including Vero-E6,LLCMK2,BHK-21,N2a,MDCK,and RfKT cell,derived from diverse mammals,i.e.monkey,mice,canine and bat,which revealed the potential of PMRV-GCCDC7 to infect a variety of hosts.Infection of BALB/c mice with PMRVGCCDC7 via intranasal inoculation led to relative weight loss,lung tissue damage and inflammation with the increase of virus titer,but no serious respiratory symptoms and death occurred.The characterization of the new reovirus from a plateau-based wild animal has expanded our knowledge of the host range of MRV and provided insight into its risk of trans-species transmission and zoonotic diseases.展开更多
An ongoing multi-country outbreak of monkeypox was reported in May 2022 with several deaths,affecting 107 countries of all six World Health Organization(WHO)regions.The WHO has declared the current monkeypox outbreak ...An ongoing multi-country outbreak of monkeypox was reported in May 2022 with several deaths,affecting 107 countries of all six World Health Organization(WHO)regions.The WHO has declared the current monkeypox outbreak to be a Public Health Emergency of International Concern.It is,thus,necessary to rapidly and accurately detect and distinguish different monkeypox virus(MPXV)clades.We designed primers and probes based on the alignment of 138 complete genomes of poxviruses.In Panel 1,we mixed one pair of primers and three probes to detect and differentiate the MPXV Western Africa(IIa,IIb clade)and Congo Basin(I clade)and other orthopoxviruses.In Panel 2,we mixed one pair of primers and two probes to detect the 2022 MPXV(B.1 lineage and its descendant lineages).In addition,we tested the specificity and sensitivity of the assay using real-time PCR.In Panel 1,the assay reproducibly identified various concentrations of two plasmids of the monkeypox virus,whereas other orthopoxviruses did not cross-react.In Panel 2,the probe annealed well to MPXV B.1 and showed the expected linearity.These two multiple real-time assays are inclusive and highly specific for identifying different clades of MPXV.展开更多
Mink has been identified as an animal with susceptibility to SARS-CoV-2 and also as the only animal with evidence to transmit the virus back to humans.Thus,the surveillance of viruses among high-density farmed minks h...Mink has been identified as an animal with susceptibility to SARS-CoV-2 and also as the only animal with evidence to transmit the virus back to humans.Thus,the surveillance of viruses among high-density farmed minks has a significant meaning for the control of zoonotic emerging diseases in humans.Within anal swabs of minks that died of unknown causes in a mink farm,mink calicivirus(MCV)and mammalian orthoreovirus(MRV)were detected and simultaneously observed within MDCK cell culture from the sample of the same lethal mink.The parallel isolation was successfully performed by utilizing cell lines from different host sources with distinct viral sensitivities,i.e.Mv.1.Lu and Vero-E6 and the two viruses were independently separated.The prevalence of the virus among the minks and its genomic characteristics were investigated through deep sequencing technology.Phylogenetic analysis of the viral genome showed a close relationship of the newly isolated MCV-GCCDC8-2020 with MCV strains belonging to the genus Vesivirus,but with unique mutations derived from the major structural protein(VP1).The reovirus MRV-GCCDC9-2020 isolated from the same mink belongs to serotype 3 mammalian orthoreovirus and genome analysis showed a potential reassortment derived from reoviruses in different species.This study provides a beneficial reference on viral co-infection within disease investigation in farmed minks and raises the concern for the virus surveillance among the high-density fed animal farms.展开更多
Background:Utilizing population-based survey data in epidemiological research with a spatial perspective can integrate valuable context into the dynamics of HIV prevalence in West Africa.However,the situation in the M...Background:Utilizing population-based survey data in epidemiological research with a spatial perspective can integrate valuable context into the dynamics of HIV prevalence in West Africa.However,the situation in the Mano River Union(MRU)countries is largely unknown.This research aims to perform an ecological study to determine the HIV prevalence patterns in MRU.Methods:We analyzed Demographic and Health Survey(DHS)and AIDS Indicator Survey(AIS)data on HIV prevalence in MRU from 2005 to 2020.We examined the country-specifc,regional-specifc and sex-specifc ratios of respondents to profle the spatial–temporal heterogeneity of HIV prevalence and determine HIV hot spots.We employed Geodetector to measure the spatial stratifed heterogeneity(SSH)of HIV prevalence for adult women and men.We assessed the comprehensive correct knowledge(CCK)about HIV/AIDS and HIV testing uptake by employing the Least Absolute Shrinkage and Selection Operator(LASSO)regression to predict which combinations of CCKs can scale up the ratio of HIV testing uptake with sex-specifc needs.Results:In our analysis,we leveraged data for 158,408 respondents from 11 surveys in the MRU.From 2005–2015,Cote d’Ivoire was the hot spot for HIV prevalence with a Gi_Bin score of 3,Z-Score 8.0–10.1 and P<0.001.From 2016 to 2020,Guinea and Sierra Leone were hot spots for HIV prevalence with a Gi_Bin score of 2,Z-Score of 3.17 and P<0.01.The SSH confrmed the signifcant diferences in HIV prevalence at the national level strata,with a higher level for Cote d’Ivoire compared to other countries in both sexes with q-values of 0.61 and 0.40,respectively.Our LASSO model predicted diferent combinations of CCKs with sex-specifc needs to improve HIV testing uptake.Conclusions:The spatial distribution of HIV prevalence in the MRU is skewed and the CCK about HIV/AIDS and HIV testing uptake are far below the threshold target set by UNAIDS for ending the epidemic in the sub-region.Geodetector detected statistically signifcant SSH within and between countries in the MRU.Our LASSO model predicted that diferent emphases should be implemented when popularizing the CCK about HIV/AIDS for adult women and men.展开更多
The vaccinia virus Tiantan(VTT)is widely utilized as a smallpox vaccine in China and holds significant impor-tance in the prevention of diseases stemming from poxvirus infections.Nevertheless,few studies have investi-...The vaccinia virus Tiantan(VTT)is widely utilized as a smallpox vaccine in China and holds significant impor-tance in the prevention of diseases stemming from poxvirus infections.Nevertheless,few studies have investi-gated the influence of VTT infection on host gene expression.In this study,we constructed time series transcriptomic profiles of HeLa cells infected with both VTT and western reserve(WR)strains.We observed similar patterns of viral gene expression,while the expression levels of host genes varied between the two strains.There was an immediate and significant repression of host gene expression,particularly in genes asso-ciated with oxidative phosphorylation.Conversely,genes involved in nerve growth factor(NGF)-stimulated transcription were significantly activated.The upregulation of genes linked to the ribonucleic acid(RNA)-induced silencing complex(RISC)suggested a potential role for posttranscriptional regulation in the interac-tion between the vaccinia virus and the host.In the later stages of infection,pathways such as extracellular matrix organization,neutrophil degranulation,complement and interferon responses,translation,and pro-grammed cell death are largely inhibited.A significant number of host genes exhibit correlations with changes in the expression levels of viral genes.The host genes that are negatively correlated with viral genes are mainly enriched in pathways associated with translation and the response to viral infection.This study significantly contributes to advancing our understanding of the dynamics between the vaccinia virus and the host,improv-ing the application of VTTs and facilitating the development of effective vaccines against diseases such as smallpox and monkeypox.展开更多
基金supported by the Natural Science Foundation of Beijing(7254390)the Youth Science Foundation of Chinese Center for Disease Control and Prevention(2024A103)to W.C.C,the National Key ResearchDevelopment Program of China(2022YFC2304100,2023YFD1800405).
文摘Ectromelia virus(ECTV),a member of the Orthopoxvirus genus,serves as both a causative agent of mousepox and a pivotal surrogate model for studying highly pathogenic orthopoxviruses.Although genomic data on ECTV remains limited,we report the isolation and characterization of a novel strain,ECTV-C-Tan-GD01,obtained from rodents in Guangdong Province,China.Nanopore sequencing yielded a complete genome(199 annotated genes,including one gene truncated at the C-terminus)with inverted terminal repeats(ITRs)harboring a conserved hairpin structure.Notably,a frameshift-inducing“G”deletion in the EV159 gene resulted in the truncation of a semaphorin-like protein.In vitro assays demonstrated cell-associated viral replication kinetics,with maximum titers achieved earlier in Vero/HeLa cells(72 h)than in BHK-21/CEF cells(84 h).Murine challenge experiments revealed extreme virulence(LD50<1 plaque-forming unit(PFU)via intranasal/footpad routes)and hepatosplenic tropism.Furthermore,ECTV-C-Tan-GD01 exhibited utility in evaluating orthopoxvirus countermeasures:a single dose of vaccinia virus Tiantan(VTT)or non-replicating vaccinia virus Tiantan(NTV)conferred cross-protection,while tecovirimat(ST-246),cidofovir(CDV),and brincidofovir(initially CMX001)significantly reduced viral loads and pathology.This study establishes ECTV-C-Tan-GD01 as a dual-purpose resource for probing orthopoxvirus evolution and advancing therapeutic development.
基金the National Key Research and Development Program of China(2021YFC0863400,2022YFC2604105).
文摘Emerging and re-emerging viruses from wild animals have seriously threatened the health of humans and domesticated animals in recent years.Herein,we isolated a new mammalian orthoreovirus(MRV),Pika/MRV/GCCDC7/2019(PMRV-GCCDC7),in the Qinghai-Tibet Plateau wild pika(Ochotona curzoniae).Though the PMRVGCCDC7 shows features of a typical reovirus with ten gene segments arranged in 3:3:4 in length,the virus belongs to an independent evolutionary branch compared to other MRVs based on phylogenetic tree analysis.The results of cellular susceptibility,species tropism,and replication kinetics of PMRV-GCCDC7 indicated the virus could infect four human cell lines(A549,Huh7,HCT,and LoVo)and six non-human cell lines,including Vero-E6,LLCMK2,BHK-21,N2a,MDCK,and RfKT cell,derived from diverse mammals,i.e.monkey,mice,canine and bat,which revealed the potential of PMRV-GCCDC7 to infect a variety of hosts.Infection of BALB/c mice with PMRVGCCDC7 via intranasal inoculation led to relative weight loss,lung tissue damage and inflammation with the increase of virus titer,but no serious respiratory symptoms and death occurred.The characterization of the new reovirus from a plateau-based wild animal has expanded our knowledge of the host range of MRV and provided insight into its risk of trans-species transmission and zoonotic diseases.
基金supported by the National Key Research and Development Program of China(2016YFD0500301,2021YFC0863300).
文摘An ongoing multi-country outbreak of monkeypox was reported in May 2022 with several deaths,affecting 107 countries of all six World Health Organization(WHO)regions.The WHO has declared the current monkeypox outbreak to be a Public Health Emergency of International Concern.It is,thus,necessary to rapidly and accurately detect and distinguish different monkeypox virus(MPXV)clades.We designed primers and probes based on the alignment of 138 complete genomes of poxviruses.In Panel 1,we mixed one pair of primers and three probes to detect and differentiate the MPXV Western Africa(IIa,IIb clade)and Congo Basin(I clade)and other orthopoxviruses.In Panel 2,we mixed one pair of primers and two probes to detect the 2022 MPXV(B.1 lineage and its descendant lineages).In addition,we tested the specificity and sensitivity of the assay using real-time PCR.In Panel 1,the assay reproducibly identified various concentrations of two plasmids of the monkeypox virus,whereas other orthopoxviruses did not cross-react.In Panel 2,the probe annealed well to MPXV B.1 and showed the expected linearity.These two multiple real-time assays are inclusive and highly specific for identifying different clades of MPXV.
基金supported in part by the Major Special Projects for Infectious Disease Research of China(Grant 2018ZX10101002-005)the Research Units of Adaptive Evolution and Control of Emerging Viruses,Chinese Academy of Medical Sciences(Grant 2018RU009)+1 种基金the National Key Research and Development Program of China(Grant 2020YFC0840800)the Young Scientists Research Fund of Academician Yunde Hou.
文摘Mink has been identified as an animal with susceptibility to SARS-CoV-2 and also as the only animal with evidence to transmit the virus back to humans.Thus,the surveillance of viruses among high-density farmed minks has a significant meaning for the control of zoonotic emerging diseases in humans.Within anal swabs of minks that died of unknown causes in a mink farm,mink calicivirus(MCV)and mammalian orthoreovirus(MRV)were detected and simultaneously observed within MDCK cell culture from the sample of the same lethal mink.The parallel isolation was successfully performed by utilizing cell lines from different host sources with distinct viral sensitivities,i.e.Mv.1.Lu and Vero-E6 and the two viruses were independently separated.The prevalence of the virus among the minks and its genomic characteristics were investigated through deep sequencing technology.Phylogenetic analysis of the viral genome showed a close relationship of the newly isolated MCV-GCCDC8-2020 with MCV strains belonging to the genus Vesivirus,but with unique mutations derived from the major structural protein(VP1).The reovirus MRV-GCCDC9-2020 isolated from the same mink belongs to serotype 3 mammalian orthoreovirus and genome analysis showed a potential reassortment derived from reoviruses in different species.This study provides a beneficial reference on viral co-infection within disease investigation in farmed minks and raises the concern for the virus surveillance among the high-density fed animal farms.
文摘Background:Utilizing population-based survey data in epidemiological research with a spatial perspective can integrate valuable context into the dynamics of HIV prevalence in West Africa.However,the situation in the Mano River Union(MRU)countries is largely unknown.This research aims to perform an ecological study to determine the HIV prevalence patterns in MRU.Methods:We analyzed Demographic and Health Survey(DHS)and AIDS Indicator Survey(AIS)data on HIV prevalence in MRU from 2005 to 2020.We examined the country-specifc,regional-specifc and sex-specifc ratios of respondents to profle the spatial–temporal heterogeneity of HIV prevalence and determine HIV hot spots.We employed Geodetector to measure the spatial stratifed heterogeneity(SSH)of HIV prevalence for adult women and men.We assessed the comprehensive correct knowledge(CCK)about HIV/AIDS and HIV testing uptake by employing the Least Absolute Shrinkage and Selection Operator(LASSO)regression to predict which combinations of CCKs can scale up the ratio of HIV testing uptake with sex-specifc needs.Results:In our analysis,we leveraged data for 158,408 respondents from 11 surveys in the MRU.From 2005–2015,Cote d’Ivoire was the hot spot for HIV prevalence with a Gi_Bin score of 3,Z-Score 8.0–10.1 and P<0.001.From 2016 to 2020,Guinea and Sierra Leone were hot spots for HIV prevalence with a Gi_Bin score of 2,Z-Score of 3.17 and P<0.01.The SSH confrmed the signifcant diferences in HIV prevalence at the national level strata,with a higher level for Cote d’Ivoire compared to other countries in both sexes with q-values of 0.61 and 0.40,respectively.Our LASSO model predicted diferent combinations of CCKs with sex-specifc needs to improve HIV testing uptake.Conclusions:The spatial distribution of HIV prevalence in the MRU is skewed and the CCK about HIV/AIDS and HIV testing uptake are far below the threshold target set by UNAIDS for ending the epidemic in the sub-region.Geodetector detected statistically signifcant SSH within and between countries in the MRU.Our LASSO model predicted that diferent emphases should be implemented when popularizing the CCK about HIV/AIDS for adult women and men.
基金supported by the National Key Research and Development Program of China(2022YFC2303401)the National Natural Science Foundation of China(82241066).
文摘The vaccinia virus Tiantan(VTT)is widely utilized as a smallpox vaccine in China and holds significant impor-tance in the prevention of diseases stemming from poxvirus infections.Nevertheless,few studies have investi-gated the influence of VTT infection on host gene expression.In this study,we constructed time series transcriptomic profiles of HeLa cells infected with both VTT and western reserve(WR)strains.We observed similar patterns of viral gene expression,while the expression levels of host genes varied between the two strains.There was an immediate and significant repression of host gene expression,particularly in genes asso-ciated with oxidative phosphorylation.Conversely,genes involved in nerve growth factor(NGF)-stimulated transcription were significantly activated.The upregulation of genes linked to the ribonucleic acid(RNA)-induced silencing complex(RISC)suggested a potential role for posttranscriptional regulation in the interac-tion between the vaccinia virus and the host.In the later stages of infection,pathways such as extracellular matrix organization,neutrophil degranulation,complement and interferon responses,translation,and pro-grammed cell death are largely inhibited.A significant number of host genes exhibit correlations with changes in the expression levels of viral genes.The host genes that are negatively correlated with viral genes are mainly enriched in pathways associated with translation and the response to viral infection.This study significantly contributes to advancing our understanding of the dynamics between the vaccinia virus and the host,improv-ing the application of VTTs and facilitating the development of effective vaccines against diseases such as smallpox and monkeypox.
